【摘要】 将克隆的鸡α干扰素（ChIFNα）基因亚克隆至酵母表达载体pPICZα-A中,构建重组质粒pPIC-ChIFNα,线性化后电转化整合到毕赤酵母菌株X-33基因组中,得到阳性菌株。经1%甲醇连续诱导4 d,SDS-PAGE检测结果表明鸡α干扰素在酵母中获得了分泌型表达,表达产物分子质量约为18 ku。细胞病变抑制法检测其效价为1×10^7.69U/mL;在鸡胚中能有效抑制禽流感病毒H9N2的增殖。针对Ⅰ群4型禽腺病毒YBAV-4株强毒的动物攻毒保护试验结果显示,表达的蛋白具有较好的抗病毒活性。更多还原

【Abstract】 The gene of chicken interferon alpha（ChIFN-α） was was subcloned into the expression vector pPICZα-A.After linearized by SacⅠ,the plasmid was transferred into yeast Pichia pastoris strain X-33.The recombinant strain was identified by PCR.The recombinant strain was induced by 1% methanol for 4 days,recombinant ChIFN-α was detected by SDS-PAGE.The assay results showed that the protein was approximately 18 ku in size,and the recombinant protein was secreted effectively in Pichia pastoris.Antiviral activities of the recombinant ChIFN-α were detected by cell pathological effect inhibition assay.The result showed the titer was about 1×10^7.69U/mL.The expressed protein could inhibit H9N2 avian influenza virus replication in chick embryo.Animal experiment suggests that recombinant protein is of good antivirus activities.更多还原